Drosophila melanogaster Ovary Dissection: A Technique for Ex Vivo Tissue Preparation
Transcript
– In order to isolate ovaries from live Drosophila flies, begin by moving an anesthetized female to one of three dissection wells filled with pre-warmed insect dissection medium. The medium will help maintain tissue integrity and viability.
Under a dissection microscope, separate the abdomen from the thorax. Then, move the abdomen into a new well. To isolate the ovaries, hold the abdomen at the posterior end and carefully scrape out the reproductive organs. Lastly, transfer the ovaries into a fresh well. After this step, you may either use the whole ovary or further dissect it into its component structures.
Each Drosophila ovary is composed of several ovarioles, consisting of a chain of egg chambers with increasing levels of maturity. To free the ovarioles, pin down the ovary at the opaque posterior end and carefully tease the protective sheath from around the ovary until ovarioles are properly separated.
In the example protocol, we will show the dissection of Drosophila ovaries for their use in live and fixed tissue microscopy.
– After sorting out five anesthetized female flies according to the text protocol, under the dissection microscope, use two pairs of forceps to sever the thorax from the abdomen. Then, carefully transfer the abdomens to the second well of the dish. Using one pair of forceps to hold the abdomen at the posterior end, use the other pair of forceps to slowly push the ovaries out.
Then, use the forceps to hold an individual ovary by the opaque posterior end and carefully move it to the third well of the dish for teasing or fixing. Carefully tease the protective sheath from around the ovaries by using a pair of forceps to hold the posterior end and lightly sweeping a teasing needle from the posterior to the anterior end.
